Isolation, characterization and biological significance of hepatitis B virus mutants from serum of a patient with immunologically negative HBV infection.

Abstract:

BACKGROUND/AIM:A low-titered hepatitis B virus infection without immunological markers was identified in a prospective study in the serum of a kidney transplant recipient by PCR. The aim of this study was to analyze HBV genomes and their biological significance. METHODS:The genome was amplified in two overlapping fragments A and B. Sequencing of 22 clones of the A- and 12 clones of the B-fragment revealed a heterogeneous virus population. A consensus and a mutant sequence were computed, representing the complete sequence of the virus population. The two sequences were compared with 41 published genomes of the different HBV geno- and serotypes. RESULTS:Ninety-five point mutations and two deletions were identified. Two mutations were observed in all clones and 17 other mutations in three or more clones. The deletions were found in ten and seven of 22 clones. They were located in the C-gene and led to stop codons yielding truncated e- and/or core proteins. In vitro transfection of DNA constructs containing these deletions demonstrated a stop of HBV replication and of HBeAg expression. Cotransfection experiments demonstrated a dominant negative effect of the mutants containing the deletions. In addition, we describe new variants of naturally occurring HBsAg mutants that may cause HBV infection less detectable by standard HBsAg measurement assays. They were characterized by two point mutations which were observed in 9 of 12 and 13 of 22 clones of the S-gene. They significantly reduced the HBsAg expression in in vitro transfection experiments. CONCLUSION:We found a patient with low-titered HBV infection, with mutations of the 'a' epitope of the Santigen as well as with mutations leading to truncated core proteins which may cause a dominant negative effect.

journal_name

J Hepatol

journal_title

Journal of hepatology

authors

Schories M,Peters T,Rasenack J

doi

10.1016/s0168-8278(00)80313-x

subject

Has Abstract

pub_date

2000-11-01 00:00:00

pages

799-811

issue

5

eissn

0168-8278

issn

1600-0641

pii

S0168-8278(00)80313-X

journal_volume

33

pub_type

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